Chapter 6 Diversity analysis
6.1 Alpha diversity
# Calculate Hill numbers
richness <- genome_counts_filt %>%
column_to_rownames(var = "genome") %>%
dplyr::select(where(~ !all(. == 0))) %>%
hilldiv(., q = 0) %>%
t() %>%
as.data.frame() %>%
dplyr::rename(richness = 1) %>%
rownames_to_column(var = "sample")
neutral <- genome_counts_filt %>%
column_to_rownames(var = "genome") %>%
dplyr::select(where(~ !all(. == 0))) %>%
hilldiv(., q = 1) %>%
t() %>%
as.data.frame() %>%
dplyr::rename(neutral = 1) %>%
rownames_to_column(var = "sample")
phylogenetic <- genome_counts_filt %>%
column_to_rownames(var = "genome") %>%
dplyr::select(where(~ !all(. == 0))) %>%
hilldiv(., q = 1, tree = genome_tree) %>%
t() %>%
as.data.frame() %>%
dplyr::rename(phylogenetic = 1) %>%
rownames_to_column(var = "sample")
# Aggregate basal GIFT into elements
dist <- genome_gifts %>%
to.elements(., GIFT_db3) %>%
traits2dist(., method = "gower")
functional <- genome_counts_filt %>%
column_to_rownames(var = "genome") %>%
dplyr::select(where(~ !all(. == 0))) %>%
hilldiv(., q = 1, dist = dist) %>%
t() %>%
as.data.frame() %>%
dplyr::rename(functional = 1) %>%
rownames_to_column(var = "sample") %>%
mutate(functional = if_else(is.nan(functional), 1, functional))
# Merge all metrics
alpha_div <- richness %>%
full_join(neutral, by = join_by(sample == sample)) %>%
full_join(phylogenetic, by = join_by(sample == sample)) %>%
full_join(functional, by = join_by(sample == sample))6.1.1 Wild samples
alpha_div %>%
pivot_longer(-sample, names_to = "metric", values_to = "value") %>%
left_join(., sample_metadata, by = join_by(sample == Tube_code)) %>%
filter(time_point=="0_Wild") %>%
mutate(metric=factor(metric,levels=c("richness","neutral","phylogenetic","functional"))) %>%
ggplot(aes(y = value, x = Population, group=Population, color=Population, fill=Population)) +
geom_boxplot(outlier.shape = NA) +
geom_jitter(alpha=0.5) +
scale_color_manual(name="Population",
breaks=c("Cold_wet","Hot_dry"),
labels=c("Cold","Hot"),
values=c('#008080', "#d57d2c")) +
scale_fill_manual(name="Population",
breaks=c("Cold_wet","Hot_dry"),
labels=c("Cold","Hot"),
values=c('#00808050', "#d57d2c50")) +
facet_wrap(. ~ metric,scales = "free") +
coord_cartesian(xlim = c(1, NA)) +
stat_compare_means(size=3, label.x=.58) +
theme_classic() +
theme(
strip.background = element_blank(),
panel.grid.minor.x = element_line(size = .1, color = "grey"),
axis.title.x = element_blank(),
axis.title.y = element_text(size=10),
axis.text.x = element_text(angle = 45, hjust = 1),
# Increase plot size
plot.title = element_text(size = 10),
axis.text = element_text(size = 8),
axis.title = element_text(size = 8)
) +
ylab("Alpha diversity")
6.1.2 Acclimation samples
alpha_div %>%
pivot_longer(-sample, names_to = "metric", values_to = "value") %>%
left_join(., sample_metadata, by = join_by(sample == Tube_code)) %>%
filter(time_point=="1_Acclimation") %>%
mutate(metric=factor(metric,levels=c("richness","neutral","phylogenetic","functional"))) %>%
ggplot(aes(y = value, x = Population, group=Population, color=Population, fill=Population)) +
geom_boxplot(outlier.shape = NA) +
geom_jitter(alpha=0.5) +
scale_color_manual(name="Population",
breaks=c("Cold_wet","Hot_dry"),
labels=c("Cold","Hot"),
values=c('#008080', "#d57d2c")) +
scale_fill_manual(name="Population",
breaks=c("Cold_wet","Hot_dry"),
labels=c("Cold","Hot"),
values=c('#00808050', "#d57d2c50")) +
facet_wrap(. ~ metric,scales = "free") +
coord_cartesian(xlim = c(1, NA)) +
stat_compare_means(size=3, label.x=.58) +
theme_classic() +
theme(
strip.background = element_blank(),
panel.grid.minor.x = element_line(size = .1, color = "grey"),
axis.title.x = element_blank(),
axis.title.y = element_text(size=10),
axis.text.x = element_text(angle = 45, hjust = 1),
# Increase plot size
plot.title = element_text(size = 10),
axis.text = element_text(size = 8),
axis.title = element_text(size = 8)
) +
ylab("Alpha diversity")
6.1.3 Antibiotics samples
alpha_div %>%
pivot_longer(-sample, names_to = "metric", values_to = "value") %>%
left_join(., sample_metadata, by = join_by(sample == Tube_code)) %>%
filter(time_point=="2_Antibiotics") %>%
mutate(metric=factor(metric,levels=c("richness","neutral","phylogenetic","functional"))) %>%
ggplot(aes(y = value, x = Population, group=Population, color=Population, fill=Population)) +
geom_boxplot(outlier.shape = NA) +
geom_jitter(alpha=0.5) +
scale_color_manual(name="Population",
breaks=c("Cold_wet","Hot_dry"),
labels=c("Cold","Hot"),
values=c('#008080', "#d57d2c")) +
scale_fill_manual(name="Population",
breaks=c("Cold_wet","Hot_dry"),
labels=c("Cold","Hot"),
values=c('#00808050', "#d57d2c50")) +
facet_wrap(. ~ metric,scales = "free") +
coord_cartesian(xlim = c(1, NA)) +
stat_compare_means(size=3, label.x=.58) +
theme_classic() +
theme(
strip.background = element_blank(),
panel.grid.minor.x = element_line(size = .1, color = "grey"),
axis.title.x = element_blank(),
axis.title.y = element_text(size=10),
axis.text.x = element_text(angle = 45, hjust = 1),
# Increase plot size
plot.title = element_text(size = 10),
axis.text = element_text(size = 8),
axis.title = element_text(size = 8)
) +
ylab("Alpha diversity")
6.1.4 Transplant_1 samples
alpha_div %>%
pivot_longer(-sample, names_to = "metric", values_to = "value") %>%
left_join(., sample_metadata, by = join_by(sample == Tube_code)) %>%
filter(time_point=="3_Transplant1") %>%
mutate(metric=factor(metric,levels=c("richness","neutral","phylogenetic","functional"))) %>%
ggplot(aes(y = value, x = type, group=type, color=type, fill=type)) +
geom_boxplot(outlier.shape = NA) +
geom_jitter(alpha=0.5) +
scale_color_manual(name="Type",
breaks=c("Control","Hot_control", "Treatment"),
labels=c("Cold-Cold","Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c", "#76b183")) +
scale_fill_manual(name="Type",
breaks=c("Control","Hot_control", "Treatment"),
labels=c("Cold-Cold","Hot-Hot", "Cold-Hot"),
values=c("#4477AA50","#d57d2c50","#76b18350")) +
facet_wrap(. ~ metric,scales = "free") +
coord_cartesian(xlim = c(1, NA)) +
stat_compare_means(size=3, label.x=.7) +
theme_classic() +
theme(
strip.background = element_blank(),
panel.grid.minor.x = element_line(size = .1, color = "grey"),
axis.title.x = element_blank(),
axis.title.y = element_text(size=10),
axis.text.x = element_text(angle = 45, hjust = 1),
# Increase plot size
plot.title = element_text(size = 10),
axis.text = element_text(size = 8),
axis.title = element_text(size = 8)
) +
ylab("Alpha diversity")
6.1.5 Transplant_2 samples
alpha_div %>%
pivot_longer(-sample, names_to = "metric", values_to = "value") %>%
left_join(., sample_metadata, by = join_by(sample == Tube_code)) %>%
filter(time_point=="4_Transplant2") %>%
mutate(metric=factor(metric,levels=c("richness","neutral","phylogenetic","functional"))) %>%
ggplot(aes(y = value, x = type, group=type, color=type, fill=type)) +
geom_boxplot(outlier.shape = NA) +
geom_jitter(alpha=0.5) +
scale_color_manual(name="Type",
breaks=c("Control","Hot_control", "Treatment"),
labels=c("Cold-Cold","Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c", "#76b183")) +
scale_fill_manual(name="Type",
breaks=c("Control","Hot_control", "Treatment"),
labels=c("Cold-Cold","Hot-Hot", "Cold-Hot"),
values=c("#4477AA50","#d57d2c50","#76b18350")) +
facet_wrap(. ~ metric,scales = "free") +
coord_cartesian(xlim = c(1, NA)) +
stat_compare_means(size=3, label.x=.7) +
theme_classic() +
theme(
strip.background = element_blank(),
panel.grid.minor.x = element_line(size = .1, color = "grey"),
axis.title.x = element_blank(),
axis.title.y = element_text(size=10),
axis.text.x = element_text(angle = 45, hjust = 1),
# Increase plot size
plot.title = element_text(size = 10),
axis.text = element_text(size = 8),
axis.title = element_text(size = 8)
) +
ylab("Alpha diversity")
6.1.6 Post-Transplant_1 samples
alpha_div %>%
pivot_longer(-sample, names_to = "metric", values_to = "value") %>%
left_join(., sample_metadata, by = join_by(sample == Tube_code)) %>%
filter(time_point=="5_Post-FMT1") %>%
mutate(metric=factor(metric,levels=c("richness","neutral","phylogenetic","functional"))) %>%
ggplot(aes(y = value, x = type, group=type, color=type, fill=type)) +
geom_boxplot(outlier.shape = NA) +
geom_jitter(alpha=0.5) +
scale_color_manual(name="Type",
breaks=c("Control","Hot_control", "Treatment"),
labels=c("Cold-Cold","Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c", "#76b183")) +
scale_fill_manual(name="Type",
breaks=c("Control","Hot_control", "Treatment"),
labels=c("Cold-Cold","Hot-Hot", "Cold-Hot"),
values=c("#4477AA50","#d57d2c50","#76b18350")) +
facet_wrap(. ~ metric,scales = "free") +
coord_cartesian(xlim = c(1, NA)) +
stat_compare_means(size=3, label.x=.7) +
theme_classic() +
theme(
strip.background = element_blank(),
panel.grid.minor.x = element_line(size = .1, color = "grey"),
axis.title.x = element_blank(),
axis.title.y = element_text(size=10),
axis.text.x = element_text(angle = 45, hjust = 1),
# Increase plot size
plot.title = element_text(size = 10),
axis.text = element_text(size = 8),
axis.title = element_text(size = 8)
) +
ylab("Alpha diversity")
6.1.7 Post-Transplant_2 samples
alpha_div %>%
pivot_longer(-sample, names_to = "metric", values_to = "value") %>%
left_join(., sample_metadata, by = join_by(sample == Tube_code)) %>%
filter(time_point=="6_Post-FMT2") %>%
mutate(metric=factor(metric,levels=c("richness","neutral","phylogenetic","functional"))) %>%
ggplot(aes(y = value, x = type, group=type, color=type, fill=type)) +
geom_boxplot(outlier.shape = NA) +
geom_jitter(alpha=0.5) +
scale_color_manual(name="Type",
breaks=c("Control","Hot_control", "Treatment"),
labels=c("Cold-control","Warm-control", "Cold-intervention"),
values=c("#4477AA","#d57d2c", "#76b183")) +
scale_fill_manual(name="Type",
breaks=c("Control","Hot_control", "Treatment"),
labels=c("Cold-control","Warm-control", "Cold-intervention"),
values=c("#4477AA50","#d57d2c50","#76b18350")) +
facet_wrap(. ~ metric,scales = "free") +
coord_cartesian(xlim = c(1, NA)) +
stat_compare_means(size=3, label.x=.7) +
theme_classic() +
theme(
strip.background = element_blank(),
panel.grid.minor.x = element_line(size = .1, color = "grey"),
axis.title.x = element_blank(),
axis.title.y = element_text(size=10),
axis.text.x = element_text(angle = 45, hjust = 1),
# Increase plot size
plot.title = element_text(size = 10),
axis.text = element_text(size = 8),
axis.title = element_text(size = 8)
) +
ylab("Alpha diversity")
6.2 Beta diversity
beta_q0n <- genome_counts_filt %>%
column_to_rownames(., "genome") %>%
hillpair(., q = 0)
beta_q1n <- genome_counts_filt %>%
column_to_rownames(., "genome") %>%
hillpair(., q = 1)
beta_q1p <- genome_counts_filt %>%
column_to_rownames(., "genome") %>%
hillpair(., q = 1, tree = genome_tree)
beta_q1f <- genome_counts_filt %>%
column_to_rownames(., "genome") %>%
hillpair(., q = 1, dist = dist)6.3 Permanovas
6.3.1 1. Are the wild populations similar?
6.3.1.1 Wild: P.muralis vs P.liolepis
wild <- meta %>%
filter(time_point == "0_Wild")
# Create a temporary modified version of genome_counts_filt
temp_genome_counts <- transform(genome_counts_filt, row.names = genome_counts_filt$genome)
temp_genome_counts$genome <- NULL
wild.counts <- temp_genome_counts[, which(colnames(temp_genome_counts) %in% rownames(wild))]
identical(sort(colnames(wild.counts)), sort(as.character(rownames(wild))))
wild_nmds <- sample_metadata %>%
filter(time_point == "0_Wild")6.3.1.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.000012 0.000012 0.0012 999 0.971
Residuals 25 0.257281 0.010291
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.973
Hot_dry 0.97302 adonis2(formula=beta_div_richness_wild$S ~ Population, data=wild[labels(beta_div_richness_wild$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 1.542719 | 0.2095041 | 6.625717 | 0.001 |
| Residual | 25 | 5.820951 | 0.7904959 | NA | NA |
| Total | 26 | 7.363669 | 1.0000000 | NA | NA |
6.3.1.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.000048 0.0000476 0.0044 999 0.938
Residuals 25 0.270114 0.0108046
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.937
Hot_dry 0.94763 adonis2(formula=beta_div_neutral_wild$S ~ Population, data=wild[labels(beta_div_neutral_wild$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 1.918266 | 0.2608511 | 8.822682 | 0.001 |
| Residual | 25 | 5.435610 | 0.7391489 | NA | NA |
| Total | 26 | 7.353876 | 1.0000000 | NA | NA |
6.3.1.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.03585 0.035847 2.4912 999 0.11
Residuals 25 0.35973 0.014389
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.11
Hot_dry 0.12705 adonis2(formula=beta_div_phylo_wild$S ~ Population, data=wild[labels(beta_div_phylo_wild$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 0.3218613 | 0.2162815 | 6.899207 | 0.001 |
| Residual | 25 | 1.1662981 | 0.7837185 | NA | NA |
| Total | 26 | 1.4881594 | 1.0000000 | NA | NA |
6.3.1.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.019387 0.019387 1.653 999 0.222
Residuals 25 0.293200 0.011728
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.213
Hot_dry 0.21033 adonis2(formula=beta_div_func_wild$S ~ Population, data=wild[labels(beta_div_func_wild$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 0.0831048 | 0.1680538 | 5.05002 | 0.048 |
| Residual | 25 | 0.4114083 | 0.8319462 | NA | NA |
| Total | 26 | 0.4945131 | 1.0000000 | NA | NA |
beta_q0n_nmds_wild <- beta_div_richness_wild$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE, trace=FALSE) %>%
vegan::scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(wild_nmds, by = join_by(sample == Tube_code))
beta_q1n_nmds_wild <- beta_div_neutral_wild$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE, trace=FALSE) %>%
vegan::scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(wild_nmds, by = join_by(sample == Tube_code))
beta_q1p_nmds_wild <- beta_div_phylo_wild$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
vegan::scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(wild_nmds, by = join_by(sample == Tube_code))
beta_q1f_nmds_wild <- beta_div_func_wild$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
vegan::scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(wild_nmds, by = join_by(sample == Tube_code))
6.3.2 2. Effect of acclimation
accli <- meta %>%
filter(time_point == "1_Acclimation")
# Create a temporary modified version of genome_counts_filt
temp_genome_counts <- transform(genome_counts_filt, row.names = genome_counts_filt$genome)
temp_genome_counts$genome <- NULL
accli.counts <- temp_genome_counts[, which(colnames(temp_genome_counts) %in% rownames(accli))]
identical(sort(colnames(accli.counts)), sort(as.character(rownames(accli))))
accli_nmds <- sample_metadata %>%
filter(time_point == "1_Acclimation")6.3.2.1 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.11796 0.117959 12.963 999 0.001 ***
Residuals 25 0.22748 0.009099
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.004
Hot_dry 0.0013711 adonis2(formula=beta_div_richness_accli$S ~ Population, data=accli[labels(beta_div_richness_accli$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 1.639807 | 0.179834 | 5.481634 | 0.001 |
| Residual | 25 | 7.478640 | 0.820166 | NA | NA |
| Total | 26 | 9.118447 | 1.000000 | NA | NA |
6.3.2.2 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.07844 0.078443 5.2384 999 0.028 *
Residuals 25 0.37437 0.014975
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.025
Hot_dry 0.030815 adonis2(formula=beta_div_neutral_accli$S ~ Population, data=accli[labels(beta_div_neutral_accli$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 1.947003 | 0.2306127 | 7.493387 | 0.001 |
| Residual | 25 | 6.495736 | 0.7693873 | NA | NA |
| Total | 26 | 8.442739 | 1.0000000 | NA | NA |
6.3.2.3 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.06739 0.067395 2.9532 999 0.089 .
Residuals 25 0.57052 0.022821
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.083
Hot_dry 0.098068 adonis2(formula=beta_div_phylo_accli$S ~ Population, data=accli[labels(beta_div_phylo_accli$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 0.2441653 | 0.1224638 | 3.488854 | 0.021 |
| Residual | 25 | 1.7496100 | 0.8775362 | NA | NA |
| Total | 26 | 1.9937754 | 1.0000000 | NA | NA |
6.3.2.4 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.02351 0.023513 0.635 999 0.442
Residuals 25 0.92569 0.037028
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.429
Hot_dry 0.43303 adonis2(formula=beta_div_func_accli$S ~ Population, data=accli[labels(beta_div_func_accli$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 0.0279416 | 0.024809 | 0.6360037 | 0.452 |
| Residual | 25 | 1.0983283 | 0.975191 | NA | NA |
| Total | 26 | 1.1262699 | 1.000000 | NA | NA |
beta_q0n_nmds_accli <- beta_div_richness_accli$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE, trace=FALSE) %>%
vegan::scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(accli_nmds, by = join_by(sample == Tube_code))
beta_q1n_nmds_accli <- beta_div_neutral_accli$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE, trace=FALSE) %>%
vegan::scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(accli_nmds, by = join_by(sample == Tube_code))
beta_q1p_nmds_accli <- beta_div_phylo_accli$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
vegan::scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(accli_nmds, by = join_by(sample == Tube_code))
beta_q1f_nmds_accli <- beta_div_func_accli$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
vegan::scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(accli_nmds, by = join_by(sample == Tube_code))
6.3.3 3. Comparison between Wild and Acclimation
accli1 <- meta %>%
filter(time_point == "0_Wild" | time_point == "1_Acclimation")
temp_genome_counts <- transform(genome_counts_filt, row.names = genome_counts_filt$genome)
temp_genome_counts$genome <- NULL
accli1.counts <- temp_genome_counts[,which(colnames(temp_genome_counts) %in% rownames(accli1))]
identical(sort(colnames(accli1.counts)),sort(as.character(rownames(accli1))))
accli1_nmds <- sample_metadata %>%
filter(time_point == "0_Wild" | time_point == "1_Acclimation")6.3.3.0.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.05014 0.050145 6.2252 999 0.022 *
Residuals 52 0.41886 0.008055
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
0_Wild 1_Acclimation
0_Wild 0.02
1_Acclimation 0.015808 adonis2(formula=beta_div_richness_accli1$S ~ time_point*Population, data=accli1[labels(beta_div_neutral_accli1$S),], permutations=999, strata=accli1$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 3.799791 | 0.222218 | 4.761789 | 0.001 |
| Residual | 50 | 13.299591 | 0.777782 | NA | NA |
| Total | 53 | 17.099381 | 1.000000 | NA | NA |
6.3.3.0.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.0199 0.0199035 2.1213 999 0.172
Residuals 52 0.4879 0.0093827
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
0_Wild 1_Acclimation
0_Wild 0.172
1_Acclimation 0.15128 adonis2(formula=beta_div_neutral_accli1$S ~ time_point*Population, data=accli1[labels(beta_div_neutral_accli1$S),], permutations=999, strata=accli1$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 4.770321 | 0.2856195 | 6.663569 | 0.001 |
| Residual | 50 | 11.931346 | 0.7143805 | NA | NA |
| Total | 53 | 16.701667 | 1.0000000 | NA | NA |
6.3.3.0.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.01334 0.013340 0.6524 999 0.441
Residuals 52 1.06332 0.020449
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
0_Wild 1_Acclimation
0_Wild 0.451
1_Acclimation 0.42294 adonis2(formula=beta_div_phylo_accli1$S ~ time_point*Population, data=accli1[labels(beta_div_phylo_accli1$S),], permutations=999, strata=accli1$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 0.855070 | 0.2267502 | 4.887385 | 0.001 |
| Residual | 50 | 2.915908 | 0.7732498 | NA | NA |
| Total | 53 | 3.770978 | 1.0000000 | NA | NA |
6.3.3.0.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.01264 0.012640 0.4951 999 0.511
Residuals 52 1.32764 0.025532
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
0_Wild 1_Acclimation
0_Wild 0.521
1_Acclimation 0.4848 adonis2(formula=beta_div_func_accli1$S ~ time_point*Population, data=accli1[labels(beta_div_func_accli1$S),], permutations=999, strata=accli1$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 0.1558147 | 0.0935514 | 1.720109 | 0.324 |
| Residual | 50 | 1.5097366 | 0.9064486 | NA | NA |
| Total | 53 | 1.6655513 | 1.0000000 | NA | NA |
beta_richness_nmds_accli1 <- beta_div_richness_accli1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(accli1_nmds, by = c("sample" = "Tube_code"))
beta_neutral_nmds_accli1 <- beta_div_neutral_accli1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(accli1_nmds, by = c("sample" = "Tube_code"))
beta_phylo_nmds_accli1 <- beta_div_phylo_accli1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(accli1_nmds, by = join_by(sample == Tube_code))
beta_func_nmds_accli1 <- beta_div_func_accli1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(accli1_nmds, by = join_by(sample == Tube_code))
6.3.4 4. Do the antibiotics work?
6.3.4.1 Antibiotics
treat1 <- meta %>%
filter(time_point == "2_Antibiotics")
temp_genome_counts <- transform(genome_counts_filt, row.names = genome_counts_filt$genome)
temp_genome_counts$genome <- NULL
treat1.counts <- temp_genome_counts[,which(colnames(temp_genome_counts) %in% rownames(treat1))]
identical(sort(colnames(treat1.counts)),sort(as.character(rownames(treat1))))
treat1_nmds <- sample_metadata %>%
filter(time_point == "2_Antibiotics")6.3.4.1.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.015319 0.0153186 6.8764 999 0.014 *
Residuals 21 0.046782 0.0022277
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.015
Hot_dry 0.015919 adonis2(formula=beta_div_richness_treat1$S ~ Population, data=treat1[labels(beta_div_neutral_treat1$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 1.356644 | 0.1527052 | 3.784762 | 0.001 |
| Residual | 21 | 7.527429 | 0.8472948 | NA | NA |
| Total | 22 | 8.884073 | 1.0000000 | NA | NA |
6.3.4.1.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.030536 0.0305358 3.8593 999 0.054 .
Residuals 21 0.166158 0.0079123
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.068
Hot_dry 0.062842 adonis2(formula=beta_div_neutral_treat1$S ~ Population, data=treat1[labels(beta_div_neutral_treat1$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 1.785669 | 0.2085055 | 5.532084 | 0.001 |
| Residual | 21 | 6.778468 | 0.7914945 | NA | NA |
| Total | 22 | 8.564137 | 1.0000000 | NA | NA |
6.3.4.1.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.012041 0.012041 0.9898 999 0.372
Residuals 21 0.255459 0.012165
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.366
Hot_dry 0.33111 adonis2(formula=beta_div_phylo_treat1$S ~ Population, data=treat1[labels(beta_div_phylo_treat1$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 0.8963254 | 0.1888758 | 4.889993 | 0.001 |
| Residual | 21 | 3.8492558 | 0.8111242 | NA | NA |
| Total | 22 | 4.7455811 | 1.0000000 | NA | NA |
6.3.4.1.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.01969 0.019691 0.4738 999 0.528
Residuals 21 0.87274 0.041559
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Cold_wet Hot_dry
Cold_wet 0.529
Hot_dry 0.49877 adonis2(formula=beta_div_func_treat1$S ~ Population, data=treat1[labels(beta_div_func_treat1$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 1 | 0.0246208 | 0.0133549 | 0.2842492 | 0.64 |
| Residual | 21 | 1.8189576 | 0.9866451 | NA | NA |
| Total | 22 | 1.8435784 | 1.0000000 | NA | NA |
beta_richness_nmds_treat1 <- beta_div_richness_treat1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(treat1_nmds, by = c("sample" = "Tube_code"))
beta_neutral_nmds_treat1 <- beta_div_neutral_treat1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(treat1_nmds, by = c("sample" = "Tube_code"))
beta_phylo_nmds_treat1 <- beta_div_phylo_treat1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(treat1_nmds, by = join_by(sample == Tube_code))
beta_func_nmds_treat1 <- beta_div_func_treat1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(treat1_nmds, by = join_by(sample == Tube_code))
6.3.4.2 Acclimation vs antibiotics
treat <- meta %>%
filter(time_point == "1_Acclimation" | time_point == "2_Antibiotics")
temp_genome_counts <- transform(genome_counts_filt, row.names = genome_counts_filt$genome)
temp_genome_counts$genome <- NULL
treat.counts <- temp_genome_counts[,which(colnames(temp_genome_counts) %in% rownames(treat))]
identical(sort(colnames(treat.counts)),sort(as.character(rownames(treat))))
treat_nmds <- sample_metadata %>%
filter(time_point == "1_Acclimation" | time_point == "2_Antibiotics")6.3.4.2.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.025318 0.0253178 6.021 999 0.011 *
Residuals 48 0.201837 0.0042049
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 2_Antibiotics
1_Acclimation 0.015
2_Antibiotics 0.017817 adonis2(formula=beta_div_richness_treat$S ~ time_point*Population, data=treat[labels(beta_div_neutral_treat$S),], permutations=999,strata=treat$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 4.885035 | 0.2455889 | 4.991572 | 0.001 |
| Residual | 46 | 15.006068 | 0.7544111 | NA | NA |
| Total | 49 | 19.891103 | 1.0000000 | NA | NA |
6.3.4.2.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.039587 0.039587 6.8387 999 0.008 **
Residuals 48 0.277854 0.005789
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 2_Antibiotics
1_Acclimation 0.008
2_Antibiotics 0.011886 adonis2(formula=beta_div_neutral_treat$S ~ time_point*Population, data=treat[labels(beta_div_neutral_treat$S),], permutations=999,strata=treat$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 5.756853 | 0.3024978 | 6.649871 | 0.001 |
| Residual | 46 | 13.274204 | 0.6975022 | NA | NA |
| Total | 49 | 19.031057 | 1.0000000 | NA | NA |
6.3.4.2.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.58372 0.58372 35.413 999 0.001 ***
Residuals 48 0.79119 0.01648
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 2_Antibiotics
1_Acclimation 0.001
2_Antibiotics 2.9795e-07 adonis2(formula=beta_div_phylo_treat$S ~ time_point*Population, data=treat[labels(beta_div_phylo_treat$S),], permutations=999,strata=treat$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 2.947011 | 0.344846 | 8.070832 | 0.001 |
| Residual | 46 | 5.598866 | 0.655154 | NA | NA |
| Total | 49 | 8.545877 | 1.000000 | NA | NA |
6.3.4.2.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.17618 0.17618 4.7941 999 0.033 *
Residuals 48 1.76400 0.03675
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 2_Antibiotics
1_Acclimation 0.026
2_Antibiotics 0.033451 adonis2(formula=beta_div_func_treat$S ~ time_point*Population, data=treat[labels(beta_div_func_treat$S),], permutations=999,strata=treat$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 1.795938 | 0.3810423 | 9.439497 | 0.001 |
| Residual | 46 | 2.917286 | 0.6189577 | NA | NA |
| Total | 49 | 4.713224 | 1.0000000 | NA | NA |
beta_richness_nmds_treat <- beta_div_richness_treat$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(treat_nmds, by = c("sample" = "Tube_code"))
beta_neutral_nmds_treat <- beta_div_neutral_treat$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(treat_nmds, by = c("sample" = "Tube_code"))
beta_phylo_nmds_treat <- beta_div_phylo_treat$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(treat_nmds, by = join_by(sample == Tube_code))
beta_func_nmds_treat <- beta_div_func_treat$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(treat_nmds, by = join_by(sample == Tube_code))
6.3.5 5. Does the FMT work?
6.3.5.1 Comparison between FMT2 vs Post-FMT2
#Create newID to identify duplicated samples
transplants_metadata<-sample_metadata%>%
mutate(Tube_code=str_remove_all(Tube_code, "_a"))
transplants_metadata$newID <- paste(transplants_metadata$Tube_code, "_", transplants_metadata$individual)
transplant3<-transplants_metadata%>%
filter(time_point == "4_Transplant2" | time_point == "6_Post-FMT2")%>%
column_to_rownames("newID")
transplant3_nmds <- transplants_metadata %>%
filter(time_point == "4_Transplant2" | time_point == "6_Post-FMT2")
full_counts<-temp_genome_counts %>%
t()%>%
as.data.frame()%>%
rownames_to_column("Tube_code")%>%
full_join(transplants_metadata,by = join_by(Tube_code == Tube_code))
transplant3_counts<-full_counts %>%
filter(time_point == "4_Transplant2" | time_point == "6_Post-FMT2") %>%
subset(select=-c(315:324)) %>%
column_to_rownames("newID")%>%
subset(select=-c(1))%>%
t() %>%
as.data.frame() %>%
mutate_if(is.character, as.numeric)
identical(sort(colnames(transplant3_counts)),sort(as.character(rownames(transplant3))))6.3.5.1.1 Number of samples used
[1] 49beta_div_richness_transplant3<-hillpair(data=transplant3_counts, q=0)
beta_div_neutral_transplant3<-hillpair(data=transplant3_counts, q=1)
beta_div_phylo_transplant3<-hillpair(data=transplant3_counts, q=1, tree=genome_tree)
beta_div_func_transplant3<-hillpair(data=transplant3_counts, q=1, dist=dist)6.3.5.1.2 Richness
adonis2(formula=beta_div_richness_transplant3$S ~ Population+time_point+type, data=transplant3[labels(beta_div_richness_transplant3$S),], permutations=999, strata=transplant3$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 3.500812 | 0.2535872 | 5.096117 | 0.001 |
| Residual | 45 | 10.304350 | 0.7464128 | NA | NA |
| Total | 48 | 13.805162 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_richness_transplant3$S,transplant3_arrange$type, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control vs Treatment | 1 | 1.4169018 | 5.739828 | 0.15622903 | 0.001 | 0.003 | * |
| Control vs Hot_control | 1 | 2.0940966 | 8.509112 | 0.21005427 | 0.001 | 0.003 | * |
| Treatment vs Hot_control | 1 | 0.3004618 | 1.265034 | 0.04179854 | 0.177 | 0.531 |
6.3.5.1.3 Neutral
adonis2(formula=beta_div_neutral_transplant3$S ~ Population+time_point+type, data=transplant3[labels(beta_div_neutral_transplant3$S),], permutations=999, strata=transplant3$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 4.128749 | 0.3031142 | 6.524331 | 0.001 |
| Residual | 45 | 9.492350 | 0.6968858 | NA | NA |
| Total | 48 | 13.621099 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_neutral_transplant3$S,transplant3_arrange$type, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control vs Treatment | 1 | 1.8758788 | 8.282671 | 0.21084796 | 0.001 | 0.003 | * |
| Control vs Hot_control | 1 | 2.4396317 | 10.635546 | 0.24945256 | 0.001 | 0.003 | * |
| Treatment vs Hot_control | 1 | 0.3158428 | 1.394345 | 0.04587515 | 0.122 | 0.366 |
6.3.5.1.4 Phylogenetic
adonis2(formula=beta_div_phylo_transplant3$S ~ Population+time_point+type, data=transplant3[labels(beta_div_phylo_transplant3$S),], permutations=999, strata=transplant3$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 0.3971179 | 0.2701357 | 5.551766 | 0.001 |
| Residual | 45 | 1.0729504 | 0.7298643 | NA | NA |
| Total | 48 | 1.4700683 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_phylo_transplant3$S,transplant3_arrange$type, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control vs Treatment | 1 | 0.14387705 | 5.735321 | 0.15612552 | 0.001 | 0.003 | * |
| Control vs Hot_control | 1 | 0.22715701 | 9.044894 | 0.22036587 | 0.001 | 0.003 | * |
| Treatment vs Hot_control | 1 | 0.04648319 | 1.704277 | 0.05550617 | 0.125 | 0.375 |
6.3.5.1.5 Functional
adonis2(formula=beta_div_func_transplant3$S ~ Population+time_point+type, data=transplant3[labels(beta_div_func_transplant3$S),], permutations=999, strata=transplant3$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 0.0880056 | 0.0736928 | 1.193332 | 0.465 |
| Residual | 45 | 1.1062168 | 0.9263072 | NA | NA |
| Total | 48 | 1.1942224 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_func_transplant3$S,transplant3_arrange$type, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control vs Treatment | 1 | 0.08177408 | 4.84137651 | 0.135077862 | 0.067 | 0.201 | |
| Control vs Hot_control | 1 | 0.05266301 | 2.16167342 | 0.063277738 | 0.199 | 0.597 | |
| Treatment vs Hot_control | 1 | -0.00189892 | -0.06088838 | -0.002104017 | 0.845 | 1.000 |
beta_richness_nmds_transplant3 <- beta_div_richness_transplant3$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(transplant3_nmds, by = join_by(sample == newID))
beta_neutral_nmds_transplant3 <- beta_div_neutral_transplant3$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(transplant3_nmds, by = join_by(sample == newID))
beta_phylo_nmds_transplant3 <- beta_div_phylo_transplant3$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(transplant3_nmds, by = join_by(sample == newID))
beta_func_nmds_transplant3 <- beta_div_func_transplant3$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(transplant3_nmds, by = join_by(sample == newID))p0<-beta_richness_nmds_transplant3 %>%
group_by(individual) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Richness beta diversity") +
theme_classic() +
theme(legend.position="none")
p1<-beta_neutral_nmds_transplant3 %>%
group_by(individual) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Neutral beta diversity") +
theme_classic() +
theme(legend.position="none")
p2<-beta_phylo_nmds_transplant3 %>%
group_by(individual) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Phylogenetic beta diversity") +
theme_classic() +
theme(legend.position="none")
p3<-beta_func_nmds_transplant3 %>%
group_by(individual) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Functional beta diversity") +
theme_classic()+
theme(legend.position="none")
6.3.5.2 Comparison between the different experimental time points (Acclimation vs Transplant samples)
The estimated time for calculating the 5151 pairwise combinations is 52 seconds.
6.3.5.3 Comparison of acclimation samples to transplant samples
transplant7<-transplants_metadata%>%
filter(time_point == "4_Transplant2" | time_point == "1_Acclimation"| time_point == "3_Transplant1")%>%
column_to_rownames("newID")
transplant7_nmds <- transplants_metadata %>%
filter(time_point == "4_Transplant2" | time_point == "1_Acclimation"| time_point == "3_Transplant1")
transplant7_counts<-full_counts %>%
filter(time_point == "4_Transplant2" | time_point == "1_Acclimation"| time_point == "3_Transplant1") %>%
subset(select=-c(315:324)) %>%
column_to_rownames("newID")%>%
subset(select=-c(1))%>%
t() %>%
as.data.frame() %>%
mutate_if(is.character, as.numeric)
transplant7_counts <- transplant7_counts[, !names(transplant7_counts) %in% c("AD45 _ LI1_2nd_2", "AD48 _ LI1_2nd_6")]
identical(sort(colnames(transplant7_counts)),sort(as.character(rownames(transplant7))))[1] TRUE6.3.5.3.1 Number of samples used
[1] 73beta_div_richness_transplant7<-hillpair(data=transplant7_counts, q=0)
beta_div_neutral_transplant7<-hillpair(data=transplant7_counts, q=1)
beta_div_phylo_transplant7<-hillpair(data=transplant7_counts, q=1, tree=genome_tree)
beta_div_func_transplant7<-hillpair(data=transplant7_counts, q=1, dist=dist)#Arrange of metadata dataframe
transplant7_arrange<-transplant7[labels(beta_div_neutral_transplant7$S),]
transplant7_arrange <- transplant7_arrange %>%
mutate(time_point = recode(time_point,
"3_Transplant1" = "Transplant",
"4_Transplant2" = "Transplant"))
transplant7_arrange$type_time <- interaction(transplant7_arrange$type, transplant7_arrange$time_point)6.3.5.3.2 Richness
adonis2(formula=beta_div_richness_transplant7$S ~ Population*time_point+type, data=transplant7[labels(beta_div_richness_transplant7$S),], permutations=999, strata=transplant7$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 6 | 5.309519 | 0.2518733 | 3.703392 | 0.001 |
| Residual | 66 | 15.770599 | 0.7481267 | NA | NA |
| Total | 72 | 21.080119 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_richness_transplant7$S,transplant7_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.36208146 | 1.0521088 | 0.06169963 | 0.306 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.28008774 | 4.6054436 | 0.22350616 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Control.Transplant | 1 | 0.55038651 | 2.2107376 | 0.08124505 | 0.007 | 0.105 | |
| Control.1_Acclimation vs Treatment.Transplant | 1 | 1.62289430 | 6.7106689 | 0.25123553 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Hot_control.Transplant | 1 | 1.73215888 | 7.4315069 | 0.25250175 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.36066298 | 5.0871520 | 0.24124415 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Control.Transplant | 1 | 0.52860586 | 2.1820402 | 0.08027507 | 0.002 | 0.030 | . |
| Treatment.1_Acclimation vs Treatment.Transplant | 1 | 1.76810026 | 7.5736721 | 0.27467042 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.Transplant | 1 | 1.87790626 | 8.3291875 | 0.27462613 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.Transplant | 1 | 1.75314247 | 8.7706781 | 0.25971282 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Treatment.Transplant | 1 | 0.27700454 | 1.5346880 | 0.07126586 | 0.067 | 1.000 | |
| Hot_control.1_Acclimation vs Hot_control.Transplant | 1 | 0.26448976 | 1.4916174 | 0.06349573 | 0.095 | 1.000 | |
| Control.Transplant vs Treatment.Transplant | 1 | 2.30884687 | 12.4299510 | 0.30002331 | 0.001 | 0.015 | . |
| Control.Transplant vs Hot_control.Transplant | 1 | 2.50396161 | 13.6713271 | 0.30604256 | 0.001 | 0.015 | . |
| Treatment.Transplant vs Hot_control.Transplant | 1 | 0.01688622 | 0.1023282 | 0.00392027 | 1.000 | 1.000 |
6.3.5.3.3 Neutral
adonis2(formula=beta_div_neutral_transplant7$S ~ Population+time_point*type, data=transplant7[labels(beta_div_neutral_transplant7$S),], permutations=999, strata=transplant7$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 8 | 7.284378 | 0.3492417 | 4.293351 | 0.001 |
| Residual | 64 | 13.573319 | 0.6507583 | NA | NA |
| Total | 72 | 20.857698 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_neutral_transplant7$S,transplant7_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.23160196 | 0.7712905 | 0.045988741 | 0.694 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.40153474 | 5.7562378 | 0.264578733 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Control.Transplant | 1 | 0.56111203 | 2.5583085 | 0.092832565 | 0.002 | 0.030 | . |
| Control.1_Acclimation vs Treatment.Transplant | 1 | 1.88709838 | 8.3257794 | 0.293929402 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Hot_control.Transplant | 1 | 2.02585000 | 9.2317432 | 0.295588471 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.63477039 | 6.8326887 | 0.299250291 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Control.Transplant | 1 | 0.61335323 | 2.8313912 | 0.101733730 | 0.002 | 0.030 | . |
| Treatment.1_Acclimation vs Treatment.Transplant | 1 | 2.10939140 | 9.4473664 | 0.320822116 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.Transplant | 1 | 2.24827218 | 10.3907678 | 0.320794118 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.Transplant | 1 | 1.87351542 | 10.3925002 | 0.293635661 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Treatment.Transplant | 1 | 0.34276062 | 1.9273510 | 0.087897118 | 0.044 | 0.660 | |
| Hot_control.1_Acclimation vs Hot_control.Transplant | 1 | 0.31638309 | 1.8072337 | 0.075911118 | 0.055 | 0.825 | |
| Control.Transplant vs Treatment.Transplant | 1 | 2.48701901 | 14.0199769 | 0.325894571 | 0.001 | 0.015 | . |
| Control.Transplant vs Hot_control.Transplant | 1 | 2.75304261 | 15.6912860 | 0.336064549 | 0.001 | 0.015 | . |
| Treatment.Transplant vs Hot_control.Transplant | 1 | 0.01764676 | 0.1022118 | 0.003915827 | 0.996 | 1.000 |
6.3.5.3.4 Phylogenetic
adonis2(formula=beta_div_phylo_transplant7$S ~ Population+time_point+type, data=transplant7[labels(beta_div_phylo_transplant7$S),], permutations=999, strata=transplant7$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 4 | 0.7377029 | 0.1879202 | 3.933904 | 0.022 |
| Residual | 68 | 3.1879143 | 0.8120798 | NA | NA |
| Total | 72 | 3.9256172 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_phylo_transplant7$S,transplant7_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.04186923 | 0.43916424 | 0.026714511 | 0.722 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.15609416 | 2.55468892 | 0.137684276 | 0.030 | 0.450 | |
| Control.1_Acclimation vs Control.Transplant | 1 | 0.03888650 | 0.83961027 | 0.032493148 | 0.487 | 1.000 | |
| Control.1_Acclimation vs Treatment.Transplant | 1 | 0.28946588 | 4.58406811 | 0.186464994 | 0.002 | 0.030 | . |
| Control.1_Acclimation vs Hot_control.Transplant | 1 | 0.31864880 | 5.37781508 | 0.196429666 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.23108846 | 4.05218385 | 0.202081922 | 0.002 | 0.030 | . |
| Treatment.1_Acclimation vs Control.Transplant | 1 | 0.11794420 | 2.69844074 | 0.097422117 | 0.045 | 0.675 | |
| Treatment.1_Acclimation vs Treatment.Transplant | 1 | 0.37640156 | 6.28511923 | 0.239113210 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.Transplant | 1 | 0.40433696 | 7.18306079 | 0.246138020 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.Transplant | 1 | 0.11597038 | 5.32063275 | 0.175478948 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Treatment.Transplant | 1 | 0.03673004 | 1.13023077 | 0.053488804 | 0.363 | 1.000 | |
| Hot_control.1_Acclimation vs Hot_control.Transplant | 1 | 0.04097680 | 1.30539166 | 0.056012432 | 0.264 | 1.000 | |
| Control.Transplant vs Treatment.Transplant | 1 | 0.21736741 | 7.59281199 | 0.207494630 | 0.001 | 0.015 | . |
| Control.Transplant vs Hot_control.Transplant | 1 | 0.25837791 | 9.19762187 | 0.228810100 | 0.001 | 0.015 | . |
| Treatment.Transplant vs Hot_control.Transplant | 1 | 0.00180330 | 0.04804393 | 0.001844435 | 0.969 | 1.000 |
6.3.5.3.5 Functional
adonis2(formula=beta_div_func_transplant7$S ~ Population+time_point+type, data=transplant7[labels(beta_div_func_transplant7$S),], permutations=999, strata=transplant7$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 4 | 0.5122014 | 0.2427344 | 5.449191 | 0.038 |
| Residual | 68 | 1.5979298 | 0.7572656 | NA | NA |
| Total | 72 | 2.1101312 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_func_transplant7$S,transplant7_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.0905830704 | 1.66462866 | 0.0942351347 | 0.247 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.0861813922 | 1.63467278 | 0.0926965190 | 0.238 | 1.000 | |
| Control.1_Acclimation vs Control.Transplant | 1 | 0.0706284677 | 2.02459114 | 0.0749166241 | 0.178 | 1.000 | |
| Control.1_Acclimation vs Treatment.Transplant | 1 | 0.3227173802 | 7.20965350 | 0.2649667516 | 0.005 | 0.075 | |
| Control.1_Acclimation vs Hot_control.Transplant | 1 | 0.3449345536 | 8.46661273 | 0.2778980651 | 0.005 | 0.075 | |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.0010225901 | 0.05430389 | 0.0033825127 | 0.658 | 1.000 | |
| Treatment.1_Acclimation vs Control.Transplant | 1 | -0.0046542303 | -0.35270812 | -0.0143102181 | 0.801 | 1.000 | |
| Treatment.1_Acclimation vs Treatment.Transplant | 1 | 0.0783171063 | 4.43726923 | 0.1815779491 | 0.073 | 1.000 | |
| Treatment.1_Acclimation vs Hot_control.Transplant | 1 | 0.0836921311 | 5.20043693 | 0.1911894629 | 0.060 | 0.900 | |
| Hot_control.1_Acclimation vs Control.Transplant | 1 | -0.0042700245 | -0.35258632 | -0.0143052054 | 0.813 | 1.000 | |
| Hot_control.1_Acclimation vs Treatment.Transplant | 1 | 0.0824858621 | 5.06251874 | 0.2019956092 | 0.063 | 0.945 | |
| Hot_control.1_Acclimation vs Hot_control.Transplant | 1 | 0.0887346857 | 5.97129920 | 0.2134795084 | 0.049 | 0.735 | |
| Control.Transplant vs Treatment.Transplant | 1 | 0.1927489878 | 15.76935832 | 0.3522355226 | 0.004 | 0.060 | |
| Control.Transplant vs Hot_control.Transplant | 1 | 0.2075592800 | 18.09824701 | 0.3686128958 | 0.003 | 0.045 | . |
| Treatment.Transplant vs Hot_control.Transplant | 1 | -0.0001900114 | -0.01304792 | -0.0005020952 | 0.702 | 1.000 |
beta_richness_nmds_transplant7 <- beta_div_richness_transplant7$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(transplant7_nmds, by = join_by(sample == newID))
beta_neutral_nmds_transplant7 <- beta_div_neutral_transplant7$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(transplant7_nmds, by = join_by(sample == newID))
beta_phylo_nmds_transplant7 <- beta_div_phylo_transplant7$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(transplant7_nmds, by = join_by(sample == newID))
beta_func_nmds_transplant7 <- beta_div_func_transplant7$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(transplant7_nmds, by = join_by(sample == newID))p0<-beta_richness_nmds_transplant7 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
scale_shape_manual(name="time_point",
breaks=c("1_Acclimation", "3_Transplant1", "4_Transplant2"),
labels=c("Acclimation", "Transplant", "Transplant"),
values=c("circle","square","square")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Richness beta diversity") +
theme_classic() +
theme(legend.position="none")
p1<-beta_neutral_nmds_transplant7 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
scale_shape_manual(name="time_point",
breaks=c("1_Acclimation", "3_Transplant1", "4_Transplant2"),
labels=c("Acclimation", "Transplant", "Transplant"),
values=c("circle","square","square")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Neutral beta diversity") +
theme_classic() +
theme(legend.position="none")
p2<-beta_phylo_nmds_transplant7 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
scale_shape_manual(name="time_point",
breaks=c("1_Acclimation", "3_Transplant1", "4_Transplant2"),
labels=c("Acclimation", "Transplant", "Transplant"),
values=c("circle","square","square")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Phylogenetic beta diversity") +
theme_classic() +
theme(legend.position="none")
p3<-beta_func_nmds_transplant7 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
scale_shape_manual(name="time_point",
breaks=c("1_Acclimation", "3_Transplant1", "4_Transplant2"),
labels=c("Acclimation", "Transplant", "Transplant"),
values=c("circle","square","square")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Functional beta diversity") +
theme_classic()+
theme(legend.position="none")
6.3.5.4 Comparison between Acclimation vs Post-FMT1
post3 <- meta %>%
filter(time_point == "1_Acclimation" | time_point == "5_Post-FMT1")
temp_genome_counts <- transform(genome_counts_filt, row.names = genome_counts_filt$genome)
temp_genome_counts$genome <- NULL
post3.counts <- temp_genome_counts[,which(colnames(temp_genome_counts) %in% rownames(post3))]
identical(sort(colnames(post3.counts)),sort(as.character(rownames(post3))))
post3_nmds <- sample_metadata %>%
filter(time_point == "1_Acclimation" | time_point == "5_Post-FMT1")6.3.5.4.1 Number of samples used
[1] 536.3.5.4.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.099607 0.049803 9.5441 999 0.001 ***
Residuals 50 0.260911 0.005218
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.00100000 0.894
Hot_control 0.00102653 0.001
Treatment 0.88832670 0.00010131 adonis2(formula=beta_div_richness_post3$S ~ time_point*type, data=post3[labels(beta_div_neutral_post3$S),], permutations=999,strata=post3$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 5 | 4.403362 | 0.2369995 | 2.919782 | 0.001 |
| Residual | 47 | 14.176268 | 0.7630005 | NA | NA |
| Total | 52 | 18.579631 | 1.0000000 | NA | NA |
pairwise <- pairwise.adonis(beta_div_richness_post3$S, post3_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.3620815 | 1.052109 | 0.06169963 | 0.339 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.2800877 | 4.605444 | 0.22350616 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.6845657 | 1.998114 | 0.11101796 | 0.005 | 0.075 | |
| Control.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.8437461 | 2.499232 | 0.14281954 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 1.1208022 | 3.568670 | 0.18236649 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.3606630 | 5.087152 | 0.24124415 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.7216200 | 2.172734 | 0.11956009 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.9551308 | 2.926054 | 0.16322910 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 1.2263345 | 4.039487 | 0.20157637 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.5_Post-FMT1 | 1 | 1.4319792 | 5.384836 | 0.25180628 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.8172413 | 3.194690 | 0.17558364 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 0.5796135 | 2.441615 | 0.13239702 | 0.002 | 0.030 | . |
| Control.5_Post-FMT1 vs Treatment.5_Post-FMT1 | 1 | 0.5615418 | 1.729004 | 0.10335366 | 0.016 | 0.240 | |
| Control.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.8438429 | 2.793772 | 0.14865413 | 0.002 | 0.030 | . |
| Treatment.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.3734921 | 1.268929 | 0.07799710 | 0.121 | 1.000 |
6.3.5.4.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.00945 0.0094472 1.1428 999 0.267
Residuals 51 0.42161 0.0082669
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 5_Post-FMT1
1_Acclimation 0.26
5_Post-FMT1 0.2901 adonis2(formula=beta_div_neutral_post3$S ~ time_point*type, data=post3[labels(beta_div_neutral_post3$S),], permutations=999,strata=post3$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 5 | 5.302354 | 0.3027004 | 4.080576 | 0.001 |
| Residual | 47 | 12.214484 | 0.6972996 | NA | NA |
| Total | 52 | 17.516838 | 1.0000000 | NA | NA |
pairwise <- pairwise.adonis(beta_div_neutral_post3$S, post3_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.2316020 | 0.7712905 | 0.04598874 | 0.715 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.4015347 | 5.7562378 | 0.26457873 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.8332162 | 2.9081103 | 0.15380227 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 1.1719595 | 4.0685514 | 0.21336447 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 1.4260875 | 5.2413171 | 0.24675104 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.6347704 | 6.8326887 | 0.29925029 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.9517634 | 3.3715700 | 0.17404733 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 1.3127773 | 4.6298256 | 0.23585668 | 0.002 | 0.030 | . |
| Treatment.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 1.6713369 | 6.2395460 | 0.28056085 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.5_Post-FMT1 | 1 | 1.5409781 | 6.8338056 | 0.29928456 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.9133614 | 4.0964534 | 0.21451383 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 0.6954835 | 3.2951234 | 0.17077493 | 0.001 | 0.015 | . |
| Control.5_Post-FMT1 vs Treatment.5_Post-FMT1 | 1 | 0.6051778 | 2.2508491 | 0.13047758 | 0.019 | 0.285 | |
| Control.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 1.0528902 | 4.1436369 | 0.20570451 | 0.002 | 0.030 | . |
| Treatment.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.4150076 | 1.6372683 | 0.09840968 | 0.050 | 0.750 |
6.3.5.4.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.05132 0.051320 2.6745 999 0.108
Residuals 51 0.97861 0.019189
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 5_Post-FMT1
1_Acclimation 0.099
5_Post-FMT1 0.10812 adonis2(formula=beta_div_phylo_post3$S ~ time_point*type, data=post3[labels(beta_div_phylo_post3$S),], permutations=999,strata=post3$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 5 | 0.7935087 | 0.2278753 | 2.774199 | 0.006 |
| Residual | 47 | 2.6886978 | 0.7721247 | NA | NA |
| Total | 52 | 3.4822065 | 1.0000000 | NA | NA |
pairwise <- pairwise.adonis(beta_div_phylo_post3$S, post3_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.04186923 | 0.4391642 | 0.02671451 | 0.749 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.15609416 | 2.5546889 | 0.13768428 | 0.033 | 0.495 | |
| Control.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.19193367 | 2.9749922 | 0.15678490 | 0.019 | 0.285 | |
| Control.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.14627288 | 1.7907381 | 0.10665035 | 0.164 | 1.000 | |
| Control.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 0.25061348 | 3.6146185 | 0.18428187 | 0.011 | 0.165 | |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.23108846 | 4.0521838 | 0.20208192 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.26358465 | 4.3608960 | 0.21417997 | 0.003 | 0.045 | . |
| Treatment.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.25319427 | 3.2738422 | 0.17915456 | 0.043 | 0.645 | |
| Treatment.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 0.39050120 | 5.9837393 | 0.27218933 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.14203376 | 5.4200212 | 0.25303529 | 0.002 | 0.030 | . |
| Hot_control.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.09666753 | 2.3682173 | 0.13635351 | 0.012 | 0.180 | |
| Hot_control.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 0.09252600 | 2.9824958 | 0.15711821 | 0.009 | 0.135 | |
| Control.5_Post-FMT1 vs Treatment.5_Post-FMT1 | 1 | 0.01842535 | 0.4144162 | 0.02688498 | 0.784 | 1.000 | |
| Control.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.05987967 | 1.7387847 | 0.09802164 | 0.105 | 1.000 | |
| Treatment.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.03212966 | 0.6477782 | 0.04139746 | 0.733 | 1.000 |
6.3.5.4.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.00554 0.0055401 0.2063 999 0.629
Residuals 51 1.36938 0.0268505
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 5_Post-FMT1
1_Acclimation 0.628
5_Post-FMT1 0.65159 adonis2(formula=beta_div_func_post3$S ~ time_point*type, data=post3[labels(beta_div_func_post3$S),], permutations=999,strata=post3$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 5 | 0.2935075 | 0.1510442 | 1.672425 | 0.054 |
| Residual | 47 | 1.6496826 | 0.8489558 | NA | NA |
| Total | 52 | 1.9431901 | 1.0000000 | NA | NA |
pairwise <- pairwise.adonis(beta_div_func_post3$S, post3_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.090583070 | 1.66462866 | 0.094235135 | 0.197 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.086181392 | 1.63467278 | 0.092696519 | 0.213 | 1.000 | |
| Control.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.028641941 | 0.50417680 | 0.030548437 | 0.553 | 1.000 | |
| Control.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.234795406 | 4.03037749 | 0.211786524 | 0.047 | 0.705 | |
| Control.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 0.134726259 | 2.20299547 | 0.121023788 | 0.171 | 1.000 | |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.001022590 | 0.05430389 | 0.003382513 | 0.658 | 1.000 | |
| Treatment.1_Acclimation vs Control.5_Post-FMT1 | 1 | 0.002157067 | 0.09411569 | 0.005847832 | 0.602 | 1.000 | |
| Treatment.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.056602363 | 2.56037069 | 0.145803909 | 0.185 | 1.000 | |
| Treatment.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 0.009569124 | 0.35095521 | 0.021463896 | 0.502 | 1.000 | |
| Hot_control.1_Acclimation vs Control.5_Post-FMT1 | 1 | -0.001745663 | -0.08225018 | -0.005167199 | 0.724 | 1.000 | |
| Hot_control.1_Acclimation vs Treatment.5_Post-FMT1 | 1 | 0.057758674 | 2.84545622 | 0.159449901 | 0.162 | 1.000 | |
| Hot_control.1_Acclimation vs Hot_control.5_Post-FMT1 | 1 | 0.005575266 | 0.21803560 | 0.013444020 | 0.541 | 1.000 | |
| Control.5_Post-FMT1 vs Treatment.5_Post-FMT1 | 1 | 0.119540855 | 4.84764704 | 0.244242909 | 0.075 | 1.000 | |
| Control.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.052587837 | 1.77308932 | 0.099762584 | 0.246 | 1.000 | |
| Treatment.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.012980354 | 0.44307662 | 0.028690955 | 0.469 | 1.000 |
beta_richness_nmds_post3 <- beta_div_richness_post3$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post3_nmds, by = c("sample" = "Tube_code"))
beta_neutral_nmds_post3 <- beta_div_neutral_post3$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post3_nmds, by = c("sample" = "Tube_code"))
beta_phylo_nmds_post3 <- beta_div_phylo_post3$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post3_nmds, by = join_by(sample == Tube_code))
beta_func_nmds_post3 <- beta_div_func_post3$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post3_nmds, by = join_by(sample == Tube_code))
6.3.5.5 Comparison between Acclimation vs Post-FMT2
post4 <- meta %>%
filter(time_point == "1_Acclimation" | time_point == "6_Post-FMT2")
temp_genome_counts <- transform(genome_counts_filt, row.names = genome_counts_filt$genome)
temp_genome_counts$genome <- NULL
post4.counts <- temp_genome_counts[,which(colnames(temp_genome_counts) %in% rownames(post4))]
identical(sort(colnames(post4.counts)),sort(as.character(rownames(post4))))
post4_nmds <- sample_metadata %>%
filter(time_point == "1_Acclimation" | time_point == "6_Post-FMT2")6.3.5.5.1 Number of samples used
[1] 546.3.5.5.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.06809 0.034047 3.8471 999 0.031 *
Residuals 51 0.45135 0.008850
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.0360000 0.891
Hot_control 0.0349385 0.003
Treatment 0.8855174 0.0047257 adonis2(formula=beta_div_richness_post4$S ~ time_point*Population, data=post4[labels(beta_div_neutral_post4$S),], permutations=999,strata=post4$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 3.310172 | 0.1883377 | 3.867324 | 0.001 |
| Residual | 50 | 14.265560 | 0.8116623 | NA | NA |
| Total | 53 | 17.575732 | 1.0000000 | NA | NA |
pairwise <- pairwise.adonis(beta_div_richness_post4$S, post4_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.3620815 | 1.052109 | 0.06169963 | 0.305 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.2800877 | 4.605444 | 0.22350616 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.8430295 | 2.845779 | 0.15100353 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.5232174 | 1.683240 | 0.09518843 | 0.022 | 0.330 | |
| Control.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 1.1217138 | 3.634271 | 0.18509835 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.3606630 | 5.087152 | 0.24124415 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.9130048 | 3.195028 | 0.16645080 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.5959230 | 1.984036 | 0.11032208 | 0.002 | 0.030 | . |
| Treatment.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 1.2747787 | 4.275366 | 0.21086503 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.6397330 | 2.913695 | 0.15405213 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.6_Post-FMT2 | 1 | 1.4575447 | 6.224524 | 0.28007456 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 0.3276169 | 1.412318 | 0.08111028 | 0.040 | 0.600 | |
| Treatment.6_Post-FMT2 vs Control.6_Post-FMT2 | 1 | 0.6463814 | 2.560441 | 0.13795154 | 0.001 | 0.015 | . |
| Treatment.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.4796256 | 1.916520 | 0.10696943 | 0.002 | 0.030 | . |
| Control.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 1.1305044 | 4.268317 | 0.21059061 | 0.001 | 0.015 | . |
6.3.5.5.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.01544 0.0154447 2.0972 999 0.163
Residuals 52 0.38294 0.0073643
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 6_Post-FMT2
1_Acclimation 0.161
6_Post-FMT2 0.15357 adonis2(formula=beta_div_neutral_post4$S ~ time_point*Population, data=post4[labels(beta_div_neutral_post4$S),], permutations=999,strata=post4$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 3.863228 | 0.229321 | 4.959284 | 0.001 |
| Residual | 50 | 12.983151 | 0.770679 | NA | NA |
| Total | 53 | 16.846379 | 1.000000 | NA | NA |
pairwise <- pairwise.adonis(beta_div_neutral_post4$S, post4_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.2316020 | 0.7712905 | 0.04598874 | 0.724 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.4015347 | 5.7562378 | 0.26457873 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 1.1746426 | 4.5564741 | 0.22165640 | 0.001 | 0.015 | . |
| Control.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.5286441 | 1.9819408 | 0.11021840 | 0.005 | 0.075 | |
| Control.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 1.3443224 | 4.9104417 | 0.23483204 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 1.6347704 | 6.8326887 | 0.29925029 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 1.3540292 | 5.3398081 | 0.25022756 | 0.002 | 0.030 | . |
| Treatment.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.6311089 | 2.4041625 | 0.13063146 | 0.002 | 0.030 | . |
| Treatment.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 1.6125755 | 5.9825981 | 0.27215155 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.6202327 | 3.1519868 | 0.16457754 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.6_Post-FMT2 | 1 | 1.5701179 | 7.6327037 | 0.32297209 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 0.3634438 | 1.7083388 | 0.09647087 | 0.031 | 0.465 | |
| Treatment.6_Post-FMT2 vs Control.6_Post-FMT2 | 1 | 1.0227481 | 4.6483346 | 0.22511910 | 0.001 | 0.015 | . |
| Treatment.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.5010202 | 2.2065321 | 0.12119453 | 0.001 | 0.015 | . |
| Control.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 1.3619424 | 5.7710313 | 0.26507845 | 0.001 | 0.015 | . |
6.3.5.5.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.06978 0.069777 5.0345 999 0.028 *
Residuals 52 0.72071 0.013860
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 6_Post-FMT2
1_Acclimation 0.024
6_Post-FMT2 0.029131 adonis2(formula=beta_div_phylo_post4$S ~ time_point*Population, data=post4[labels(beta_div_phylo_post4$S),], permutations=999,strata=post4$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 0.757493 | 0.2376349 | 5.195124 | 0.001 |
| Residual | 50 | 2.430141 | 0.7623651 | NA | NA |
| Total | 53 | 3.187634 | 1.0000000 | NA | NA |
pairwise <- pairwise.adonis(beta_div_phylo_post4$S, post4_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.04186923 | 0.4391642 | 0.02671451 | 0.735 | 1.000 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.15609416 | 2.5546889 | 0.13768428 | 0.038 | 0.570 | |
| Control.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.26322331 | 4.3060281 | 0.21205664 | 0.007 | 0.105 | |
| Control.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.16047895 | 2.5405742 | 0.13702781 | 0.037 | 0.555 | |
| Control.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 0.25529510 | 4.0109138 | 0.20043631 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.23108846 | 4.0521838 | 0.20208192 | 0.005 | 0.075 | |
| Treatment.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.36496892 | 6.3966666 | 0.28560797 | 0.001 | 0.015 | . |
| Treatment.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.22628210 | 3.8292220 | 0.19311005 | 0.014 | 0.210 | |
| Treatment.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 0.34830814 | 5.8463335 | 0.26761166 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.10002871 | 4.3836237 | 0.21505615 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.12577510 | 5.0601287 | 0.24027055 | 0.001 | 0.015 | . |
| Hot_control.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 0.06334378 | 2.4997737 | 0.13512455 | 0.018 | 0.270 | |
| Treatment.6_Post-FMT2 vs Control.6_Post-FMT2 | 1 | 0.05927454 | 2.3820253 | 0.12958449 | 0.027 | 0.405 | |
| Treatment.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.06906280 | 2.7224602 | 0.14541146 | 0.007 | 0.105 | |
| Control.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.11081709 | 4.0436561 | 0.20174244 | 0.002 | 0.030 | . |
6.3.5.5.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 1 0.00527 0.005269 0.1889 999 0.665
Residuals 52 1.45058 0.027896
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
1_Acclimation 6_Post-FMT2
1_Acclimation 0.657
6_Post-FMT2 0.66565 adonis2(formula=beta_div_func_post4$S ~ time_point*Population, data=post4[labels(beta_div_func_post4$S),], permutations=999,strata=post4$individual) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 3 | 0.0773959 | 0.0417692 | 0.726498 | 0.31 |
| Residual | 50 | 1.7755477 | 0.9582308 | NA | NA |
| Total | 53 | 1.8529436 | 1.0000000 | NA | NA |
pairwise <- pairwise.adonis(beta_div_func_post4$S, post4_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.1_Acclimation vs Treatment.1_Acclimation | 1 | 0.0905830704 | 1.664628661 | 0.0942351347 | 0.222 | 1 | |
| Control.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.0861813922 | 1.634672780 | 0.0926965190 | 0.253 | 1 | |
| Control.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.1197900330 | 2.213130846 | 0.1215129274 | 0.164 | 1 | |
| Control.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.1125623700 | 2.150784454 | 0.1184953995 | 0.179 | 1 | |
| Control.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 0.0657004998 | 0.954588109 | 0.0563026423 | 0.284 | 1 | |
| Treatment.1_Acclimation vs Hot_control.1_Acclimation | 1 | 0.0010225901 | 0.054303886 | 0.0033825127 | 0.634 | 1 | |
| Treatment.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | -0.0005177706 | -0.025585400 | -0.0016016487 | 0.710 | 1 | |
| Treatment.1_Acclimation vs Control.6_Post-FMT2 | 1 | 0.0013301207 | 0.072110871 | 0.0044867082 | 0.609 | 1 | |
| Treatment.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 0.0060959077 | 0.174487757 | 0.0107878382 | 0.575 | 1 | |
| Hot_control.1_Acclimation vs Treatment.6_Post-FMT2 | 1 | 0.0010345754 | 0.055797964 | 0.0034752533 | 0.654 | 1 | |
| Hot_control.1_Acclimation vs Control.6_Post-FMT2 | 1 | -0.0001056284 | -0.006306177 | -0.0003942915 | 0.679 | 1 | |
| Hot_control.1_Acclimation vs Hot_control.6_Post-FMT2 | 1 | 0.0017235602 | 0.051851181 | 0.0032302306 | 0.744 | 1 | |
| Treatment.6_Post-FMT2 vs Control.6_Post-FMT2 | 1 | -0.0080428882 | -0.442986255 | -0.0284750185 | 0.847 | 1 | |
| Treatment.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | -0.0011796256 | -0.034047378 | -0.0021324990 | 0.902 | 1 | |
| Control.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.0036300838 | 0.110487573 | 0.0068581148 | 0.673 | 1 |
beta_richness_nmds_post4 <- beta_div_richness_post4$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post4_nmds, by = c("sample" = "Tube_code"))
beta_neutral_nmds_post4 <- beta_div_neutral_post4$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post4_nmds, by = c("sample" = "Tube_code"))
beta_phylo_nmds_post4 <- beta_div_phylo_post4$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post4_nmds, by = join_by(sample == Tube_code))
beta_func_nmds_post4 <- beta_div_func_post4$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post4_nmds, by = join_by(sample == Tube_code))
6.3.6 6. Are there differences between the control and the treatment group?
6.3.6.1 After 1 week –> Post-FMT1
post1 <- meta %>%
filter(time_point == "5_Post-FMT1")
post1.counts <- temp_genome_counts[,which(colnames(temp_genome_counts) %in% rownames(post1))]
identical(sort(colnames(post1.counts)),sort(as.character(rownames(post1))))
post1_nmds <- sample_metadata %>%
filter(time_point == "5_Post-FMT1")6.3.6.1.1 Number of samples used
[1] 266.3.6.1.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.017675 0.0088373 2.3825 999 0.111
Residuals 23 0.085312 0.0037092
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.0050000 0.682
Hot_control 0.0068795 0.229
Treatment 0.6248469 0.2084296 adonis2(formula=beta_div_richness_post1$S ~ Population+type, data=post1[labels(beta_div_richness_post1$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 1.195567 | 0.1448246 | 1.947534 | 0.001 |
| Residual | 23 | 7.059710 | 0.8551754 | NA | NA |
| Total | 25 | 8.255277 | 1.0000000 | NA | NA |
pairs Df SumsOfSqs F.Model R2 p.value p.adjusted sig
1 Control vs Treatment 1 0.5615418 1.729004 0.1033537 0.013 0.039 .
2 Control vs Hot_control 1 0.8438429 2.793772 0.1486541 0.001 0.003 *
3 Treatment vs Hot_control 1 0.3734921 1.268929 0.0779971 0.106 0.318 6.3.6.1.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.011001 0.0055005 0.6303 999 0.575
Residuals 23 0.200714 0.0087267
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.23900 0.966
Hot_control 0.21166 0.451
Treatment 0.95468 0.43604 adonis2(formula=beta_div_neutral_post1$S ~ Population+type, data=post1[labels(beta_div_neutral_post1$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 1.395968 | 0.1900228 | 2.697931 | 0.001 |
| Residual | 23 | 5.950350 | 0.8099772 | NA | NA |
| Total | 25 | 7.346318 | 1.0000000 | NA | NA |
pairs Df SumsOfSqs F.Model R2 p.value p.adjusted sig
1 Control vs Treatment 1 0.6051778 2.250849 0.13047758 0.010 0.030 .
2 Control vs Hot_control 1 1.0528902 4.143637 0.20570451 0.002 0.006 *
3 Treatment vs Hot_control 1 0.4150076 1.637268 0.09840968 0.059 0.177 6.3.6.1.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.00440 0.0021994 0.1369 999 0.9
Residuals 23 0.36941 0.0160614
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.92000 0.702
Hot_control 0.91505 0.758
Treatment 0.63312 0.73046 adonis2(formula=beta_div_phylo_post1$S ~ Population+type, data=post1[labels(beta_div_phylo_post1$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 0.0745104 | 0.0705947 | 0.8735033 | 0.532 |
| Residual | 23 | 0.9809570 | 0.9294053 | NA | NA |
| Total | 25 | 1.0554673 | 1.0000000 | NA | NA |
pairs Df SumsOfSqs F.Model R2 p.value p.adjusted sig
1 Control vs Treatment 1 0.01842535 0.4144162 0.02688498 0.798 1.000
2 Control vs Hot_control 1 0.05987967 1.7387847 0.09802164 0.119 0.357
3 Treatment vs Hot_control 1 0.03212966 0.6477782 0.04139746 0.681 1.000 6.3.6.1.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.00400 0.0019999 0.1431 999 0.862
Residuals 23 0.32135 0.0139717
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.61000 0.707
Hot_control 0.60188 0.848
Treatment 0.74597 0.84473 adonis2(formula=beta_div_func_post1$S ~ Population+type, data=post1[labels(beta_div_func_post1$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 0.1230554 | 0.1608583 | 2.204479 | 0.16 |
| Residual | 23 | 0.6419374 | 0.8391417 | NA | NA |
| Total | 25 | 0.7649929 | 1.0000000 | NA | NA |
pairs Df SumsOfSqs F.Model R2 p.value p.adjusted sig
1 Control vs Treatment 1 0.11954085 4.8476470 0.24424291 0.070 0.210
2 Control vs Hot_control 1 0.05258784 1.7730893 0.09976258 0.232 0.696
3 Treatment vs Hot_control 1 0.01298035 0.4430766 0.02869096 0.473 1.000 beta_richness_nmds_post1 <- beta_div_richness_post1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post1_nmds, by = join_by(sample == Tube_code))
beta_neutral_nmds_post1 <- beta_div_neutral_post1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post1_nmds, by = join_by(sample == Tube_code))
beta_phylogenetic_nmds_post1 <- beta_div_phylo_post1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post1_nmds, by = join_by(sample == Tube_code))
beta_functional_nmds_post1 <- beta_div_func_post1$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post1_nmds, by = join_by(sample == Tube_code))p0<-beta_richness_nmds_post1 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Richness beta diversity") +
theme_classic() +
theme(legend.position="none")
p1<-beta_neutral_nmds_post1 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Neutral beta diversity") +
theme_classic() +
theme(legend.position="none")
p2<-beta_phylogenetic_nmds_post1 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Phylogenetic beta diversity") +
theme_classic() +
theme(legend.position="none")
p3<-beta_functional_nmds_post1 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Functional beta diversity") +
theme_classic()+
theme(legend.position="none")
6.3.6.2 After 2 weeks –>Post-FMT2
post2 <- meta %>%
filter(time_point == "6_Post-FMT2")
post2.counts <- temp_genome_counts[,which(colnames(temp_genome_counts) %in% rownames(post2))]
identical(sort(colnames(post2.counts)),sort(as.character(rownames(post2))))
post2_nmds <- sample_metadata %>%
filter(time_point == "6_Post-FMT2")6.3.6.2.1 Number of samples used
[1] 276.3.6.2.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.002011 0.0010056 0.1982 999 0.832
Residuals 24 0.121775 0.0050740
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.69800 0.814
Hot_control 0.67789 0.649
Treatment 0.79246 0.59820 adonis2(formula=beta_div_richness_post2$S ~ type, data=post2[labels(beta_div_richness_post2$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 1.504341 | 0.1967776 | 2.939822 | 0.001 |
| Residual | 24 | 6.140538 | 0.8032224 | NA | NA |
| Total | 26 | 7.644879 | 1.0000000 | NA | NA |
| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Treatment vs Control | 1 | 0.6463814 | 2.560441 | 0.1379515 | 0.001 | 0.003 | * |
| Treatment vs Hot_control | 1 | 0.4796256 | 1.916520 | 0.1069694 | 0.001 | 0.003 | * |
| Control vs Hot_control | 1 | 1.1305044 | 4.268317 | 0.2105906 | 0.001 | 0.003 | * |
6.3.6.2.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.008262 0.0041311 0.8024 999 0.492
Residuals 24 0.123559 0.0051483
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.47700 0.655
Hot_control 0.44675 0.267
Treatment 0.65989 0.25095 adonis2(formula=beta_div_neutral_post2$S ~ type, data=post2[labels(beta_div_neutral_post2$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 1.923807 | 0.2603795 | 4.224537 | 0.001 |
| Residual | 24 | 5.464666 | 0.7396205 | NA | NA |
| Total | 26 | 7.388473 | 1.0000000 | NA | NA |
| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Treatment vs Control | 1 | 1.0227481 | 4.648335 | 0.2251191 | 0.001 | 0.003 | * |
| Treatment vs Hot_control | 1 | 0.5010202 | 2.206532 | 0.1211945 | 0.002 | 0.006 | * |
| Control vs Hot_control | 1 | 1.3619424 | 5.771031 | 0.2650785 | 0.001 | 0.003 | * |
6.3.6.2.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.000407 0.0002034 0.0487 999 0.954
Residuals 24 0.100305 0.0041794
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.93000 0.849
Hot_control 0.93765 0.761
Treatment 0.83933 0.76015 adonis2(formula=beta_div_phylo_post2$S ~ type, data=post2[labels(beta_div_phylo_post2$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 0.1594363 | 0.2042241 | 3.079623 | 0.001 |
| Residual | 24 | 0.6212564 | 0.7957759 | NA | NA |
| Total | 26 | 0.7806927 | 1.0000000 | NA | NA |
| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Treatment vs Control | 1 | 0.05927454 | 2.382025 | 0.1295845 | 0.034 | 0.102 | |
| Treatment vs Hot_control | 1 | 0.06906280 | 2.722460 | 0.1454115 | 0.004 | 0.012 | . |
| Control vs Hot_control | 1 | 0.11081709 | 4.043656 | 0.2017424 | 0.001 | 0.003 | * |
6.3.6.2.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.01259 0.0062962 0.3249 999 0.798
Residuals 24 0.46507 0.0193778
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.54400 0.653
Hot_control 0.45381 0.798
Treatment 0.57452 0.74365 adonis2(formula=beta_div_func_post2$S ~ type, data=post2[labels(beta_div_func_post2$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | -0.0037283 | -0.0054704 | -0.065288 | 0.915 |
| Residual | 24 | 0.6852623 | 1.0054704 | NA | NA |
| Total | 26 | 0.6815340 | 1.0000000 | NA | NA |
| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Treatment vs Control | 1 | -0.008042888 | -0.44298625 | -0.028475019 | 0.842 | 1 | |
| Treatment vs Hot_control | 1 | -0.001179626 | -0.03404738 | -0.002132499 | 0.902 | 1 | |
| Control vs Hot_control | 1 | 0.003630084 | 0.11048757 | 0.006858115 | 0.681 | 1 |
beta_richness_nmds_post2 <- beta_div_richness_post2$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post2_nmds, by = join_by(sample == Tube_code))
beta_neutral_nmds_post2 <- beta_div_neutral_post2$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post2_nmds, by = join_by(sample == Tube_code))
beta_phylogenetic_nmds_post2 <- beta_div_phylo_post2$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post2_nmds, by = join_by(sample == Tube_code))
beta_functional_nmds_post2 <- beta_div_func_post2$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post2_nmds, by = join_by(sample == Tube_code))p0<-beta_richness_nmds_post2 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Richness beta diversity") +
theme_classic() +
theme(legend.position="none")
p1<-beta_neutral_nmds_post2 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Neutral beta diversity") +
theme_classic() +
theme(legend.position="none")
p2<-beta_phylogenetic_nmds_post2 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Phylogenetic beta diversity") +
theme_classic() +
theme(legend.position="none")
p3<-beta_functional_nmds_post2 %>%
group_by(type) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Functional beta diversity") +
theme_classic()+
theme(legend.position="none")
6.3.6.3 Post1 vs Post2
post5 <- meta %>%
filter(time_point == "6_Post-FMT2" | time_point == "5_Post-FMT1")
post5.counts <- temp_genome_counts[,which(colnames(temp_genome_counts) %in% rownames(post5))]
identical(sort(colnames(post5.counts)),sort(as.character(rownames(post5))))
post5_nmds <- sample_metadata %>%
filter(time_point == "6_Post-FMT2"| time_point == "5_Post-FMT1")6.3.6.3.1 Number of samples used
[1] 536.3.6.3.2 Richness
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.01841 0.0092048 1.7364 999 0.187
Residuals 50 0.26505 0.0053010
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.045000 0.699
Hot_control 0.039117 0.202
Treatment 0.716358 0.218648 adonis2(formula=beta_div_richness_post5$S ~ type, data=post5[labels(beta_div_richness_post5$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 2.28826 | 0.1390012 | 4.036044 | 0.001 |
| Residual | 50 | 14.17390 | 0.8609988 | NA | NA |
| Total | 52 | 16.46216 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_richness_post5$S,post5_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.5_Post-FMT1 vs Treatment.5_Post-FMT1 | 1 | 0.5615418 | 1.729004 | 0.10335366 | 0.020 | 0.300 | |
| Control.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.8438429 | 2.793772 | 0.14865413 | 0.001 | 0.015 | . |
| Control.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.7628135 | 2.683925 | 0.14364890 | 0.002 | 0.030 | . |
| Control.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.3432605 | 1.148733 | 0.06698647 | 0.258 | 1.000 | |
| Control.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 1.1269580 | 3.799256 | 0.19188884 | 0.001 | 0.015 | . |
| Treatment.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.3734921 | 1.268929 | 0.07799710 | 0.118 | 1.000 | |
| Treatment.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.3571397 | 1.297184 | 0.07959561 | 0.119 | 1.000 | |
| Treatment.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.7769467 | 2.670898 | 0.15114670 | 0.001 | 0.015 | . |
| Treatment.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 0.6502360 | 2.253407 | 0.13060650 | 0.001 | 0.015 | . |
| Hot_control.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.4132091 | 1.616138 | 0.09174188 | 0.009 | 0.135 | |
| Hot_control.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 1.0163992 | 3.760571 | 0.19030682 | 0.001 | 0.015 | . |
| Hot_control.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 0.2732563 | 1.019281 | 0.05988979 | 0.436 | 1.000 | |
| Treatment.6_Post-FMT2 vs Control.6_Post-FMT2 | 1 | 0.6463814 | 2.560441 | 0.13795154 | 0.001 | 0.015 | . |
| Treatment.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.4796256 | 1.916520 | 0.10696943 | 0.001 | 0.015 | . |
| Control.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 1.1305044 | 4.268317 | 0.21059061 | 0.001 | 0.015 | . |
6.3.6.3.3 Neutral
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.01992 0.0099587 1.565 999 0.233
Residuals 50 0.31818 0.0063636
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.12300 0.868
Hot_control 0.10701 0.162
Treatment 0.87156 0.17449 adonis2(formula=beta_div_neutral_post5$S ~ type, data=post5[labels(beta_div_neutral_post5$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 2.928527 | 0.1906221 | 5.887921 | 0.001 |
| Residual | 50 | 12.434468 | 0.8093779 | NA | NA |
| Total | 52 | 15.362995 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_neutral_post5$S,post5_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.5_Post-FMT1 vs Treatment.5_Post-FMT1 | 1 | 0.6051778 | 2.250849 | 0.13047758 | 0.015 | 0.225 | |
| Control.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 1.0528902 | 4.143637 | 0.20570451 | 0.001 | 0.015 | . |
| Control.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.8908158 | 3.714692 | 0.18842252 | 0.001 | 0.015 | . |
| Control.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.3860927 | 1.552176 | 0.08843210 | 0.082 | 1.000 | |
| Control.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 1.3122237 | 5.130273 | 0.24279254 | 0.001 | 0.015 | . |
| Treatment.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.4150076 | 1.637268 | 0.09840968 | 0.039 | 0.585 | |
| Treatment.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.3157079 | 1.325203 | 0.08117526 | 0.175 | 1.000 | |
| Treatment.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 1.0579520 | 4.270010 | 0.22158835 | 0.001 | 0.015 | . |
| Treatment.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 0.7454015 | 2.920049 | 0.16294873 | 0.001 | 0.015 | . |
| Hot_control.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.4377161 | 1.942126 | 0.10824392 | 0.009 | 0.135 | |
| Hot_control.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 1.3766597 | 5.875279 | 0.26858075 | 0.001 | 0.015 | . |
| Hot_control.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 0.3176516 | 1.316137 | 0.07600637 | 0.167 | 1.000 | |
| Treatment.6_Post-FMT2 vs Control.6_Post-FMT2 | 1 | 1.0227481 | 4.648335 | 0.22511910 | 0.002 | 0.030 | . |
| Treatment.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.5010202 | 2.206532 | 0.12119453 | 0.001 | 0.015 | . |
| Control.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 1.3619424 | 5.771031 | 0.26507845 | 0.001 | 0.015 | . |
6.3.6.3.4 Phylogenetic
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.00051 0.0002543 0.0265 999 0.972
Residuals 50 0.47996 0.0095993
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.89200 0.831
Hot_control 0.88926 0.926
Treatment 0.82391 0.91902 adonis2(formula=beta_div_phylo_post5$S ~ type, data=post5[labels(beta_div_phylo_post5$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 0.1778594 | 0.0910216 | 2.503403 | 0.007 |
| Residual | 50 | 1.7761762 | 0.9089784 | NA | NA |
| Total | 52 | 1.9540356 | 1.0000000 | NA | NA |
pairwise<-pairwise.adonis(beta_div_phylo_post5$S,post5_arrange$type_time, perm=999)
pairwise%>%
tt()| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.5_Post-FMT1 vs Treatment.5_Post-FMT1 | 1 | 0.01842535 | 0.4144162 | 0.02688498 | 0.779 | 1.000 | |
| Control.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.05987967 | 1.7387847 | 0.09802164 | 0.111 | 1.000 | |
| Control.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.07917244 | 3.0180046 | 0.15869197 | 0.009 | 0.135 | |
| Control.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.04335491 | 1.5335604 | 0.08746429 | 0.190 | 1.000 | |
| Control.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 0.10783045 | 3.7500438 | 0.18987521 | 0.002 | 0.030 | . |
| Treatment.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.03212966 | 0.6477782 | 0.04139746 | 0.681 | 1.000 | |
| Treatment.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.06393539 | 1.5651817 | 0.09448624 | 0.147 | 1.000 | |
| Treatment.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.05265949 | 1.2240203 | 0.07544494 | 0.291 | 1.000 | |
| Treatment.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 0.09753501 | 2.2402429 | 0.12994265 | 0.018 | 0.270 | |
| Hot_control.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.07228545 | 2.3279593 | 0.12701683 | 0.036 | 0.540 | |
| Hot_control.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.11759094 | 3.5538444 | 0.18174658 | 0.001 | 0.015 | . |
| Hot_control.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 0.06667255 | 1.9859527 | 0.11041687 | 0.100 | 1.000 | |
| Treatment.6_Post-FMT2 vs Control.6_Post-FMT2 | 1 | 0.05927454 | 2.3820253 | 0.12958449 | 0.020 | 0.300 | |
| Treatment.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.06906280 | 2.7224602 | 0.14541146 | 0.005 | 0.075 | |
| Control.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.11081709 | 4.0436561 | 0.20174244 | 0.001 | 0.015 | . |
6.3.6.3.5 Functional
Permutation test for homogeneity of multivariate dispersions
Permutation: free
Number of permutations: 999
Response: Distances
Df Sum Sq Mean Sq F N.Perm Pr(>F)
Groups 2 0.00785 0.0039232 0.2322 999 0.791
Residuals 50 0.84483 0.0168966
Pairwise comparisons:
(Observed p-value below diagonal, permuted p-value above diagonal)
Control Hot_control Treatment
Control 0.53800 0.605
Hot_control 0.52384 0.849
Treatment 0.58787 0.85068 adonis2(formula=beta_div_func_post5$S ~ type, data=post5[labels(beta_div_func_post5$S),], permutations=999) %>%
as.matrix() %>%
kable()| Df | SumOfSqs | R2 | F | Pr(>F) | |
|---|---|---|---|---|---|
| Model | 2 | 0.0668661 | 0.0466009 | 1.221967 | 0.315 |
| Residual | 50 | 1.3680018 | 0.9533991 | NA | NA |
| Total | 52 | 1.4348679 | 1.0000000 | NA | NA |
| pairs | Df | SumsOfSqs | F.Model | R2 | p.value | p.adjusted | sig |
|---|---|---|---|---|---|---|---|
| Control.5_Post-FMT1 vs Treatment.5_Post-FMT1 | 1 | 0.1195408549 | 4.84764704 | 0.2442429086 | 0.072 | 1 | |
| Control.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.0525878365 | 1.77308932 | 0.0997625840 | 0.245 | 1 | |
| Control.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.0265995825 | 1.17541806 | 0.0684360667 | 0.293 | 1 | |
| Control.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.0145818992 | 0.69975992 | 0.0419023938 | 0.417 | 1 | |
| Control.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | -0.0080695208 | -0.21617323 | -0.0136958691 | 0.914 | 1 | |
| Treatment.5_Post-FMT1 vs Hot_control.5_Post-FMT1 | 1 | 0.0129803540 | 0.44307662 | 0.0286909552 | 0.467 | 1 | |
| Treatment.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | 0.0267162134 | 1.22560581 | 0.0755352882 | 0.321 | 1 | |
| Treatment.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.0384388433 | 1.93281582 | 0.1141461550 | 0.217 | 1 | |
| Treatment.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | 0.0553988290 | 1.47819391 | 0.0897060633 | 0.235 | 1 | |
| Hot_control.5_Post-FMT1 vs Treatment.6_Post-FMT2 | 1 | -0.0040061386 | -0.14850469 | -0.0093684974 | 0.767 | 1 | |
| Hot_control.5_Post-FMT1 vs Control.6_Post-FMT2 | 1 | 0.0024023972 | 0.09538980 | 0.0059265296 | 0.627 | 1 | |
| Hot_control.5_Post-FMT1 vs Hot_control.6_Post-FMT2 | 1 | -0.0004960759 | -0.01190328 | -0.0007445087 | 0.838 | 1 | |
| Treatment.6_Post-FMT2 vs Control.6_Post-FMT2 | 1 | -0.0080428882 | -0.44298625 | -0.0284750185 | 0.846 | 1 | |
| Treatment.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | -0.0011796256 | -0.03404738 | -0.0021324990 | 0.921 | 1 | |
| Control.6_Post-FMT2 vs Hot_control.6_Post-FMT2 | 1 | 0.0036300838 | 0.11048757 | 0.0068581148 | 0.697 | 1 |
beta_richness_nmds_post5 <- beta_div_richness_post5$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post5_nmds, by = join_by(sample == Tube_code))
beta_neutral_nmds_post5 <- beta_div_neutral_post5$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post5_nmds, by = join_by(sample == Tube_code))
beta_phylogenetic_nmds_post5 <- beta_div_phylo_post5$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post5_nmds, by = join_by(sample == Tube_code))
beta_functional_nmds_post5 <- beta_div_func_post5$S %>%
metaMDS(.,trymax = 500, k=2, verbosity=FALSE) %>%
scores() %>%
as_tibble(., rownames = "sample") %>%
left_join(post5_nmds, by = join_by(sample == Tube_code))p0<-beta_richness_nmds_post5 %>%
group_by(type, time_point) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Richness beta diversity") +
theme_classic() +
theme(legend.position="none")
p1<-beta_neutral_nmds_post5 %>%
group_by(type, time_point) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y = "NMDS2", x="NMDS1 \n Neutral beta diversity") +
theme_classic() +
theme(legend.position="none")
p2<-beta_phylogenetic_nmds_post5 %>%
group_by(type, time_point) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Phylogenetic beta diversity") +
theme_classic() +
theme(legend.position="none")
p3<-beta_functional_nmds_post5 %>%
group_by(type, time_point) %>%
mutate(x_cen = mean(NMDS1, na.rm = TRUE)) %>%
mutate(y_cen = mean(NMDS2, na.rm = TRUE)) %>%
ungroup() %>%
ggplot(., aes(x=NMDS1,y=NMDS2, color=type, shape=time_point)) +
scale_color_manual(name="Type",
breaks=c("Control", "Hot_control", "Treatment"),
labels=c("Cold-Cold", "Hot-Hot", "Cold-Hot"),
values=c("#4477AA","#d57d2c","#76b183")) +
geom_point(size=2) +
geom_segment(aes(x=x_cen, y=y_cen, xend=NMDS1, yend=NMDS2), alpha=0.2) +
labs(y= element_blank (), x="NMDS1 \n Functional beta diversity") +
theme_classic()+
theme(legend.position="none")